The Effect And Mechanism Of N6-Methyladenosine(m~6A) In Cisplatin-induced Acute Kidney Injury

Objectives:To determine the effect and mechanism of Fat mass and obesity-associated protein(FTO)in cisplatin-induced acute kidney injury.Methods:c57BL/6 mice were randomly divided to four groups including conrol group,meclofenamic acid control group,cisplatin group and cisplatin+MA group.The latter two group got 17 mg/kg cisplatin and 3 days later the mice were sacrificed.Blood usea nitrogen,serum creatine,and histological changes were estimated.Human kidney proximal tubular cells(HK2 cells)were divided to conrol group,MA control group,cisplatin group and cisplatin+MA group,cisplatin+FTO si-RNA,cisplatin+FTO si-RNA+MA,cisplatin+pc-FTO and cisplatin+pc FTO+MA.FTO,Bcl-2,Bax,Cleaved Caspase-3,p53 expression levels were assessed by Western Blot.TUNEL was used to test apoptosis level and m~6A level was estimated by Dot Blot.p53 mRNA level was estimated by rl-qPCR.Results:Animal experiments showed that the expression of FTO protein in kidney tissue decreased and m~6A modification increased after cisplatin treatment.MA inhibited the expression of FTO protein,increased the level of RNA m~6A modification in kidney tissue of mice,aggravated the decline of renal function and pathological damage caused by cisplatin,and increased the ratio of Bax/Bcl-2,the expression of Cleaved Caspase-3 p53 and TUNEL positive rate.Cell experiments showed that the expression of FTO protein decreased and m~6A modification increased in HK2 cells treated with cisplatin.MA inhibited the expression of FTO protein in cells,increased the level of RNA m~6A modification,and increased the ratio of Bax/Bcl-2,the expression of Cleaved Caspase-3and TUNEL positive rate.The results of FTO si-RNA and chemical inhibitors were basically the same.The over-expression plasmid of FTO corrected the increase of Bax/Bcl-2 ratio,the expression of Cleaved Caspase-3,p53 and the positive rate of TUNEL caused by cisplatin and MA.Conclusions:FTO protects against cisplatin induced renal injury by regulating m~6A and p53 signaling related apoptosis.